Cytoskeletal and nuclear matrix proteins (NMPs) play an important role in the bio-chemical events involved in differentiation and transformation of mammalian cells. Certain of these markers, F/G-Actin and bladder cancer- related NMPs, provide a unique means for early identification of bladder cancer as well as assessment of field disease effects. New methods of isolating, purifying reagents and labeling these biomarkers, combined with major advances in automated image cytophotometry (both optical density and fluorescence based) provide an opportunity to develop multi-marker diagnostic assays. CytoDiagnostics Inc. (CDI), a national oncology reference laboratory, is collaborating with Matritech, Inc. to investigate the dual marker combination of Actin/NMP to identify bladder cancer cells in cytology specimens. Matritech is an expert in nuclear matrix protein chemistry, while CDI has expertise with Actin and automated image analysis. The two Companies with Oklahoma University department of urology, will investigate the utility of Actin/NMP for detection of bladder cancer, assessment of field disease dynamics, relationship to recurrence, therapeutic responsiveness, and progression. The instrumentation applied to the project will include Flow Cytometry and quantitative image cytophotometry with Zeiss-IBAS automated image system having unique software modifications.